Publications

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    ABSTRACT: Objectives the aim of this study was to evaluate the effect of SLPI on the growth and biological processes of Candida albicans. Methods Two C. albicans strains were used in this study, a clinical isolate resistant to fluconazole (PRI) and a reference strain ATCC 24433. The minimal inhibitory concentration (MIC) was determined according to the CLSI methodology. The influence of SLPI on secreted serine proteinase activities (SSP) was measured by the cleavage of specific substrate, and surface hydrophobicity was determined by the aqueous-hydrocarbon biphasic separation method. Flow cytometry was performed to investigate receptors for SLPI and variations in the cell wall mannoprotein expression. Interaction between yeast and epithelium was assessed using the MA-104 cells lineage. Ultrastructure was analyzed by transmission electron microscopy (TEM). Results MIC values were calculated as 18 and 18.9 μM for the PRI and ATCC 24433, respectively. SSP activity was reduced by 48.8% by 18 μM of SLPI and cell surface hydrophobicity increased by 11.1%. Flow cytometry suggest the existence of SLPI binding sites on the surface of the yeast. Results showed a reduction in the expression of mannoproteins in 20.8% by the cells treated with 80 μM of SLPI, and 18 μM reduced the adhesion of yeasts to mammalian cells in 60.1%. TEM revealed ultrastructural changes in cells treated with 80 μM of SLPI, such as the presence of membrane-like structures within the cytoplasm. Conclusions SLPI exerts a significant influence on C. albicans viability and biological processes. Considering its constitutive and physiologic features, SLPI may become a promising tool for the development of new methodologies for the treatment and control of candidiasis.
    Archives of Oral Biology. 01/2014;
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    ABSTRACT: Electrochemical treatment is an alternative modality for tumor treatment based on the application of a low intensity direct electric current to the tumor tissue through two or more platinum electrodes placed within the tumor zone or in the surrounding areas. This treatment is noted for its great effectiveness, minimal invasiveness and local effect. Several studies have been conducted worldwide to evaluate the antitumoral effect of this therapy. In all these studies a variety of biochemical and physiological responses of tumors to the applied treatment have been obtained. By this reason, researchers have suggested various mechanisms to explain how direct electric current destroys tumor cells. Although, it is generally accepted this treatment induces electrolysis, electroosmosis and electroporation in tumoral tissues. However, action mechanism of this alternative modality on the tumor tissue is not well understood. Although the principle of Electrochemical treatment is simple, a standardized method is not yet available. The mechanism by which Electrochemical treatment affects tumor growth and survival may represent more complex process. The present work analyzes the latest and most important research done on the electrochemical treatment of tumors. We conclude with our point of view about the destruction mechanism features of this alternative therapy. Also, we suggest some mechanisms and strategies from the thermodynamic point of view for this therapy. In the area of Electrochemical treatment of cancer this tool has been exploited very little and much work remains to be done. Electrochemical treatment constitutes a good therapeutic option for patients that have failed the conventional oncology methods.
    Chinese Journal of Cancer Research 04/2013; 25(2):223-234. · 0.45 Impact Factor
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    ABSTRACT: Influenza viruses cause highly contagious acute respiratory illnesses with significant mortality, especially among young children, elderly people, and individuals with serious medical conditions. This encourages the development of new treatments for human flu. Biotherapies are diluted solutions prepared from biological products compounded following homeopathic procedures. To develop a biotherapy prepared from the infectious influenza A virus (A/Aichi/2/68 H3N2) and to verify its in vitro response. The ultradiluted influenza virus solution was prepared in the homeopathic dilution 30dH, it was termed Influenzinum RC. The cellular alterations induced by this preparation were analyzed by optical and electron microscopy, MTT and neutral red assays. Glycolytic metabolism (PFK-1) was studied by spectrophotometric assay. Additionally, the production of tumor necrosis factor-α (TNF-α) by J774.G8 macrophage cells was quantified by ELISA before and after infection with H3N2 influenza virus and treatment. Influenzinum RC did not cause cytotoxic effects but induced morphological alterations in Madin-Darby canine kidney (MDCK) cells. After 30 days, a significant increase (p < 0.05) in mitosis rate was detected compared to control. MDCK mitochondrial activity was changed after treatment for 10 and 30 days. Treatment significantly diminished (p < 0.05) PFK-1 activity. TNF-α in biotherapy-stimulated J774.G8 macrophages indicated a significant (p < 0.05) increase in this cytokine when the cell supernatant was analyzed. Influenzinum RC altered cellular and biochemical features of MDCK and J774G8 cells.
    Homeopathy: the journal of the Faculty of Homeopathy 01/2013; 102(1):31-40. · 1.13 Impact Factor
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    ABSTRACT: This work aimed to produce poly(methyl methacrylate) nanoparticles for use in drug encapsulation. The polymer nanoparticles were produced using miniemulsion polymerization technique. Monomer miniemulsion showed moderate stability and polymer average particle size was about 90 nm. PMMA nanoparticles were tested for toxicity in human leukemic cell strain K562 and they did not show any adverse effect on cell viability. Therefore, poly(methyl methacrylate) nanoparticles are suitable to encapsulate antitumor agents.
    Macromolecular Symposia 09/2012; 319(1).
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    ABSTRACT: Staphylococcus lugdunensis is an unusually virulent coagulase-negative species, which causes serious infection similar to S. aureus. We evaluated the expression of virulence factors such as S. lugdunensis synergistic haemolysin (SLUSH), fibrinogen-binding protein (Fbl), biofilm production and biofilm-production-related genes in 23 S. lugdunensis clinical isolates and one type strain that had been previously characterized for their genotypes. In addition, the biofilm composition and the ability of isolates to adhere to and invade human epithelial lung cells were also investigated. The PCR method used detected the presence of slush and intercellular adhesin (ica) virulence genes in all isolates. All isolates produced the Fbl protein and, with the exception of the type strain, all isolates produced the SLUSH haemolysin. Fourteen (60.9 %) isolates produced biofilms. The detachment assay, using sodium metaperiodate or proteolytic enzymes to analyse the biofilm composition, showed protein-mediated biofilms in two representative isolates, one for each colony type (rough and smooth). All strongly biofilm-producing isolates, including three with rough colony morphology, had the same prevalent PFGE pattern. However, among the representative strains tested, only the S. lugdunensis isolate that formed rough colonies was able to adhere to and invade A549 cell monolayers in the same quantities as those observed with S. aureus isolates (P = 1.000). No significant adhesion or invasion was observed for the other isolates in comparison with the S. aureus isolate, independent of biofilm production or clonality. Our results could explain the incredible ability of this pathogen to cause infections that are as aggressive as S. aureus. In addition, the ability of S. lugdunensis to adhere to and invade eukaryotic cells was also noticed for isolates with rough colony morphology, reinforcing the increased virulence in this species.
    Journal of Medical Microbiology 11/2011; 61(Pt 4):463-9. · 2.30 Impact Factor
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    ABSTRACT: The literature shows that the effects of direct electric currents on biological material are numerous, including bactericidal, fungicidal, parasiticidal, and anti-tumoral, among others. Non-pathogenic trypanosomatids, such as Herpetomonas samuelpessoai, have emerged as important models for the study of basic biological processes performed by a eukaryotic cell. The present study reports a dose-dependent anti-protozoan effect of direct electric treatment with both cathodic and anodic current flows on H. samuelpessoai cells. The damaging effects can be attributable to the electrolysis products generated during electric stimulation. The pH of the cell suspension was progressively augmented from 7.4 to 10.5 after the cathodic treatment. In contrast, the anodic treatment caused a pH decrease varying from 7.4 to 6.5. Transmission electron microscopy analyses revealed profound alterations in vital cellular structures (e.g., mitochondrion, kinetoplast, flagellum, flagellar pocket, nucleus, and plasma membrane) after exposure to both cathodic and anodic current flows. Specifically, cathodic current flow treatment induced the appearance of autophagic-like structures on parasite cells, while those submitted to an anodic current flow presented marked disorganization of plasma membrane and necrotic appearance. However, parasites treated in the intermediary chamber (without contact with the electrodes) did not present significant changes in viability or morphology, and no pH variation was detected in this system. The use of H. samuelpessoai as a biological model and the direct electric current experimental approach used in our study provide important information for understanding the mechanisms involved in the cytotoxic effects of this physical agent. Bioelectromagnetics. © 2011 Wiley Periodicals, Inc.
    Bioelectromagnetics 10/2011; · 2.02 Impact Factor
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    ABSTRACT: In this study, zinc phthalocyanine (ZnPc) was loaded onto poly-ɛ-caprolactone (PCL) nanoparticles (NPs) using a solvent emulsification-evaporation method. The process yield and encapsulation efficiency were 74.2% ± 1.2% and 67.1% ± 0.9%, respectively. The NPs had a mean diameter of 187.4 ± 2.1 nm, narrow distribution size with a polydispersity index of 0.096 ± 0.004, zeta potential of -4.85 ± 0.21 mV, and spherical shape. ZnPc has sustained release, following Higuchi's kinetics. The photobiological activity of the ZnPc-loaded NPs was evaluated on human lung adenocarcinoma A549 cells. Cells were incubated with free ZnPc or ZnPc-loaded NPs for 4 h and then washed with phosphate-buffered saline. Culture medium was added to the wells containing the cells. Finally, the cells were exposed to red light (660 nm) with a light dose of 100 J/cm(2). The cellular viability was determined after 24 h of incubation. ZnPc-loaded NPs and free photosensitizer eliminated about 95.9% ± 1.8% and 28.7% ± 2.2% of A549 cells, respectively. The phototoxicity was time dependent up to 4 h and concentration dependent at 0-5 μg ZnPc. The cells viability decreased with the increase of the light dose in the range of 10-100 J/cm(2). Intense lysis was observed in the cells incubated with the ZnPcloaded NPs and irradiated with red light. ZnPc-loaded PCL NPs are the release systems that promise photodynamic therapy use.
    International Journal of Nanomedicine 01/2011; 6:227-38. · 4.20 Impact Factor
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    ABSTRACT:   There are several kinds of oral soft tissue lesions that are common manifestations observed in human immunodeficiency virus (HIV)-infected children; for example, linear gingival erythema (LGE) that is a distinctive fiery red band along the margin of the gingivae. The etiology and pathogenesis of LGE are questionable, but a candidal origin has been suggested. Proteases are key virulence attributes produced by a variety of pathogenic fungi, including Candida. The objective of the present study is to identify the protease production in Candida species including, C. albicans (n=5), C. dubliniensis (n=1) and C. tropicalis (n=1), isolated directly from typical LGE lesions observed in six HIV-positive children, and also to test the effect of a serine protease inhibitor on the interaction of Candida spp. and epithelial cells in vitro. The ability of Candida strains to release proteases in the culture supernatant fluids was visualized by gelatin-SDS-PAGE. Gel strips containing 30-fold concentrated supernatant (1.5×10(8) yeasts) were incubated at 37°C for 48 h in 50 mM sodium phosphate buffer, pH 5.5. The concentrated supernatants were also incubated with fibronectin, laminin, immunoglobulin G, bovine serum albumin and human serum albumin. The effect of serine protease inhibitor on the interaction of Candida spp. and epithelial cells (MA 104) was measured after pre-treatment of fungi with the inhibitor (phenylmethylsulphonyl fluoride, PMSF). All the extracellular proteases were completely inhibited by PMSF, identifying these activities as serine-type proteases. Interestingly, a common 62-kDa serine protease was observed in all Candida strains. The culture supernatants, rich in serine protease activities, cleaved several soluble proteinaceous substrates. Additionally, we demonstrated that pre-treatment of C. albicans, C. dubliniensis and C. tropicalis with PMSF diminished the interaction with epithelial cells. Collectively, our results show that Candida spp. isolated from LGE lesions produced and secreted serine proteases and these enzymes may be involved in the initial colonization events.
    Journal of Oral Pathology and Medicine 11/2010; 39(10):753-60. · 2.06 Impact Factor
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    ABSTRACT: This study describes the expression of acidic ectophosphatase activity on twenty isolates of C. albicans from oral cavities of HIV-infected children (HIV+) and compares them with fifteen isolates from HIV-negative children (HIV-), as well as the fungal adhesion to epithelial cells and medical records. The activities were measured in intact cells grown in BHI medium for 48 h at 37 degrees C. Phosphatase activity was assayed at pH 5.5 using 4-methylumbelliferyl phosphate. Yeast adhesion was measured using the MA 104 epithelial cell line. Mean values of ectophosphatase activity were 610.27 +/- 166.36 and 241.25 +/- 78.96 picomoles 4-methylumbelliferone/h/10(7) cells for HIV+ and HIV- group, respectively (P = 0.049). No correlation between C. albicans enzyme activity from HIV children with viral load and CD4 percentual was observed. Yeasts with high enzyme activity, isolated from HIV+ children showed greater adherence than yeasts with basal levels of ectophosphatases from HIV- (Spearman correlation, r = 0.8). Surface phosphatase activity was apparently involved in the adhesion to host cells, as the enhanced attachment of C. albicans to host epithelial cells was reversed by pretreatment of yeast with sodium orthovanadate (1 mM), an acid phosphatase inhibitor. These results show that C. albicans from HIV+ has an ectophosphatase activity significantly higher than the other isolates. Yeasts expressing higher levels of surface phosphatase activity showed greater adhesion to epithelial cells. So, the activity of acidic surface phosphatases on these cells may contribute to the early mechanisms required for disease establishment.
    Oral Diseases 03/2010; 16(5):431-7. · 2.38 Impact Factor
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    ABSTRACT: Cadmium is an important toxic environmental heavy metal. Several studies have demonstrated that a major site of cadmium toxicity in humans and in other animals is the proximal tubule of the kidney. A well established model for nefrotoxicity is the use of in vitro technique with proximal tubule epithelial cell lines, as LLC-PK1. Herein, we have the intention to study the possible protective effect of highdiluted CdCl2 solutions. In a blinding way, LLC-PK1 cells were pre-treated with highdiluted cadmium chloride in the potencies 10 cH, 15 cH and 20cH. After 4 days, these cells have received CdCl2 in a pre-determined toxic concentration. The cell viability was assessed by MTT assay. We have identified a protective effect of two CdCl2 highdiluted solutions, 10 cH and 20 cH, when cells were intoxicated by sublethal CdCl2 concentration. The results indicate that probably the highdilutions have an expressive action on cells in sublethal intoxication.
    International Journal of High Dilution Research. 01/2010;
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    ABSTRACT: Inhibition of tumor growth induced by treatment with direct electric current (DC) has been reported in several models. One of the mechanisms responsible for the antitumoral activity of DC is the generation of oxidative species, known as chloramines. With the aim of increasing chloramine production in the electrolytic medium and optimizing the antitumoral effects of DC, poly(ɛ-caprolactone) (PCL) nanoparticles (NPs) loaded with the amino acid tyrosine were obtained. The physical-chemical characterization showed that the NPs presented size in nanometric range and monomodal distribution. A slightly negative electrokinetic potential was also found in both blank NPs and L-tyrosine-loaded PCL NPs. The yield of the loading process was approximately 50%. Within 3 h of dissolution assay, a burst release of about 80% L-tyrosine was obtained. The in vitro cytotoxicity of DC was significantly increased when associated with L-tyrosine-loaded NPs, using a murine multidrug-resistant melanoma cell line model. This study showed that the use of the combination of nanotechnology and DC has a promising antineoplastic potential and opens a new perspective in cancer therapy.
    International Journal of Nanomedicine 01/2010; 5:961-71. · 4.20 Impact Factor
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    ABSTRACT: Background: although Hahnemann described the fifty-milesimal (LM) method in the 6th edition of the Organon of the Medical Art, very little research has been carried out on the physical chemical properties of these homeopathic preparations. Furthermore, there is still no evidence allowing for the correlation between the alleged physical chemical properties and the biological effects of high dilutions. Aims: to evaluate physical chemical characteristics of LM preparations including electrical conductivity, pH and refraction index, and their effect on biological experimental models. Materials and methods: preparations tested for physical chemical analysis were dilutions 1 lm to 10 lm of Euphorbia tirucalli L. prepared from the latex and the juice of the plant. To rule the seasonal characteristics of this plant, 2 different populations were used, one collected in June 2007 and the other in May 2008. Furthermore, the cytotoxic effect of Euphorbia tirucalli 5 lm was tested on human breast cancer cells (MCF7) through MTT assay. Some differences among the two collections were observed. However, any clear correlation could be observed between physical chemical properties and biological activity.
    International Journal of High Dilution Research. 01/2010;
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    ABSTRACT: In this study, oral sustained release mucoadhesive nystatin tablets were developed to increase nystatin contact time with the oral cavity and mask its unpleasant taste. The best formulation studied included sustained release agents and it was submitted to physical-mechanical characterization, taste assessment and clinical test in twelve patients. The ultraviolet-visible nystatin methodology was also developed and validated in parallel as an alternative to the pharmacopoeial microbiological dosage method. The best formulation developed in this study included sustained release agents. The efficacy of this formulation was verified through a clinical assessment, showing that this formulation is more effective (100%) than the commercial oral nystatin suspension used traditionally (50%). Moreover, the UV absorption spectrophotometry method developed to validate the methodology for nystatin content analysis for new oral tablets was shown to be specific, linear, exact and reproducible, as recommended by the ICH regulations. The oral nystatin tablets developed showed to present faster therapeutic response than the oral aqueous solution through the preliminary clinical assays. The UV absorption spectrophotometry method showed to be an attractive test for the usual routine in the pharmaceutical industry.
    Drug Development and Industrial Pharmacy 12/2009; 36(5):594-600. · 1.54 Impact Factor
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    Carla Holandino
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    ABSTRACT: RESUMO Sistemas dinamizados (SD) têm sua origem através da interação entre dois processos: diluição e agitação. Alguns autores têm sugerido que o processo de preparação é capaz de alterar as propriedades físicas e químicas da água. Entretanto, nos dias de hoje, pouco se conhece acerca dos SD, em parte porque a própria dinâmica da água, assim como suas propriedades físico-químicas, não são completamente compreendidas. O presente artigo revisa o estado atual dos conhecimentos a esse respeito e sumariza a linha de pesquisa sendo desenvolvida no Brasil, com pouco suporte em termos de recursos, pelo grupo liderado pela autora. Palavras-chave Altas diluições; Mecanismo de ação; Propriedades das altas diluições; Memória da água; Revisão Homeopathy and Experimental Models for the Understanding of the Physical-Chemical and Biological Properties of Dynamized Systems ABSTRACT Dynamized systems (DS) result from the interaction between two procedures: dilution and agitation. Some authors have suggested that the process of preparation is able to change the physical and chemical properties of water. However, in our days DS remain poorly understood, partially because dynamics of water itself as well as its physical-chemical properties are not also well known. This present paper reviews the state of knowledge in this regard and summarizes the research line followed by the group chaired by the author.
    01/2009; 723:15-18.
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    ABSTRACT: Crithidia deanei is an insect trypanosomatid that harbors a bacterial endosymbiont in its cytoplasm. In this work, we have demonstrated the influence of the endosymbiont on the interaction of C. deanei with mammalian fibroblasts, also implicating the surface leishmanolysin-like molecules of C. deanei in this process. The wild strain of C. deanei expressed a higher amount (2-fold) of leishmanolysin-like molecules in the parasite surface than the aposymbiotic strain. The treatment of parasites with anti-leishmanolysin antibodies or the fibroblasts with purified leishmanolysin-like molecules from C. deanei significantly reduced the association index. The aposymbiotic strain of C. deanei presented interaction rates about 2- and 3-fold lower with fibroblasts than the endosymbiont-bearing counterpart after 1 and 2h, respectively. However, the association indexes were similar after 3 and 4h of interaction. Additionally, we observed a 2-fold increase in the association index after 24-96 h of parasite-fibroblast interaction when compared to the interaction process performed for 4h, irrespective to the presence of the endosymbiont, suggesting that fibroblasts support multiplication and survival of C. deanei. Both parasite strains were able to induce fibroblast lysis. Interestingly, the wild strain led to a 2-fold increase in fibroblasts death in comparison to the aposymbiotic strain after 48-96 h. We also showed that both wild and aposymbiotic biotinylated live parasites recognized the same receptor in the fibroblast cells.
    Experimental Parasitology 01/2009; 121(3):246-53. · 2.15 Impact Factor
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    ABSTRACT: Plant-derived substances have been considered as important sources of drugs, including antineoplasic agents. Babassu mesocarp is popularly used in Brazil as a food additive, and in popular medicine against several conditions, such as inflammations, menstrual pains and leukaemia. From babassu Orbignya speciosa (Mart.) Barb. Rodr. [Arecaceae (Palmae)] epicarp/mesocarp, an ethanol extract was prepared and named OSEME, which was tested on the viability,morphology and metabolism of several cell lines, such as the leukaemic cell lines, HL-60, K562 and the latter multidrug resistant counterpart K562-Lucena 1, the human breast cancer cell line MCF-7, the mouse fibroblast cell line 3T3-L1 and fresh human lymphocytes. OSEME promoted a dose-dependent decrease on the viability of all cells. This effect was much more pronounced on the tumoral cell lines than on non-tumoral cells, a phenomenon revealed by the dose of OSEME which promotes half of maximal effect (ID50). The decrease on viability was followed by shrinkage of cells, alteration on their morphology, and a markedly nuclear condensation. Curiously, stimulation of 6-phosphofructokinase activity (6.6-times) was observed on HL-60 cells, treated with OSEME, when compared to control treated with ethanol (vehicle). These results support evidences to suggest OSEME as a promising source of novel antineoplasic agents.
    Anais da Academia Brasileira de Ciências 10/2008; 80(3):467-76. · 0.85 Impact Factor
  • Carla Holandino
    Homeopathy: the journal of the Faculty of Homeopathy 05/2008; 97(2):57-8. · 1.13 Impact Factor
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    ABSTRACT: The most common way to perform succussions is to place a liquid preparation inside a glass vessel and beat it vigorously against a hard elastic surface, either manually or using a mechanical apparatus. This procedure has been assumed able to transfer mechanical energy to the molecular level, where it becomes available to perform chemical work. Such interpretation has been enforced by observed changes in the electrical conductivity (EC) of High Dilutions (HD) due to succussion. In order to address this question, we compared the electrical conductivity changes of HD prepared from Vincristine sulfate (VCR) samples with those of an inert solvent. Samples were produced through manual and mechanical succusions in order to observe the influence of bubbles production. The results confirmed the timing of EC changes but these were equivalent for VCR and solvent, except for VCR 1cH samples. Also, the production of bubbles does not affect the EC in an extent able to distinguish succussion procedures. We concluded that the physical-chemical properties of HD can be modeled by chemical and diffusive mechanisms typical of distilled water. Keywords: Succussion, electrical conductivity, high dilutions, distilled water, modeling. Modelagem de propriedades físico-químicas das Altas Diluições: o estudo da condutividade elétrica Resumo A maneira mais comum de realizar sucussões é manter uma preparação líquida dentro de um frasco de vidro, e agitá-la violentamente usando um dispositivo mecânico ou batendo-a, manualmente, contra um anteparo rígido, porém elástico. Este procedimento tem sido interpretado como um mecanismo capaz de transferir a energia mecânica da agitação, para níveis moleculares, onde ela se tornaria disponível para a realização de trabalho químico. Tal interpretação tem sido estimulada pela observação de alterações na condutividade elétrica (CE) de Altas Diluições (HD), devido à sucussão. Visando modelar esta questão, comparou-se a alteração da CE de amostras preparadas a partir de Sulfato de Vincristina (VCR) com amostras equivalentes (controle) preparadas apenas com o solvente água destilada. As amostras foram produzidas por sucussão mecânica e manual, a fimd e observar a influência da produção de bolhas. Os resultados confirma que a CE varia com o tempo, porém tais variações são equivalentes para VCR e controle, exceto para as amostras VCR 1cH. Também, a produção de bolhas não afeta a CE de forma a permitir usá-la para distinguir os diferentes procedimentos de sucussão. Concluimos que as propriedades físico-químicas da HDs podem ser modeladas por mecanismos químicos e difusivos, típicos para a água destilada. Palavras-chave: Sucussão, condutividade elétrca, altas diluições, água destilada, modelagem. Modelo de propiedades físico-químicas de las Altas diluciones: un estudio de la conductividad eléctrica Resumen La manera más común para llevar a cabo sucuciones es mantener una preparación líquida dentro de un recipiente de vidrio, agitando violentamente utilizando para esto un aparato mecánico, o golpes manuales contra una superfície dura mas elástica. Este procedimiento ha sido interpretado como un mecanismo capaz de transferir la energía mecánica a nivel molecular, donde es posible realizar el trabajo químico. Tal interpretación ha sido estimulada por los cambios observados en la conductividad eléctrica (CE) de las Altas diluciones (HD), debido al procedimiento de la sucucion. A fin de abordar este punto, se compararon los cambios en la conductividad eléctrica de HD preparadas a partir de Sulfato de Vincristina (VCR) con muestras de solvente inerte. Las muestras fueron producidas a través de sucuciones mecánicas y manuales, con el fin de observar la influencia de la producción de burbujas. Los resultados confirmaron el cambio em la CE, pero estos cambios fueron equivalentes para VCR y solvente, con excepción de las muestras VCR 1cH. Asimismo, la producción de burbujas no afecta la CE en una medida capaz de distinguir las diferentes maneras de sucución. Concluimos que las propiedades físico-químicas de HD pueden ser modeladas por mecanismos químicos y difusivos típicos del agua destilada. Palabras-clave: Sucución, conductividad eléctrica, alta diluciones, agua destilada, modelo. Correspondence author: Carlos Renato Zacharias, zacha@feg.unesp.br How to cite this article: Holandino C, Harduim R, Veiga VF, Garcia, S. Zacharias CR. Modeling physical-chemical properties of High Dilutions: an electrical conductivity study. Int J High Dilution Res [online]. 2008 [cited YYYY Mmm DD]; 7(25): 165-173. Available from: http://www.feg.unesp.br/~ojs/index.php/ijhdr/article/view/319/375. La manera más común para llevar a cabo sucuciones es mantener una preparación líquida dentro de un recipiente de vidrio, agitando violentamente utilizando para esto un aparato mecánico, o golpes manuales contra una superfície dura mas elástica. Este procedimiento ha sido interpretado como un mecanismo capaz de transferir la energía mecánica a nivel molecular, donde es posible realizar el trabajo químico. Tal interpretación ha sido estimulada por los cambios observados en la conductividad eléctrica (CE) de las Altas diluciones (HD), debido al procedimiento de la sucucion. A fin de abordar este punto, se compararon los cambios en la conductividad eléctrica de HD preparadas a partir de Sulfato de Vincristina (VCR) con muestras de solvente inerte. Las muestras fueron producidas a través de sucuciones mecánicas y manuales, con el fin de observar la influencia de la producción de burbujas. Los resultados confirmaron el cambio em la CE, pero estos cambios fueron equivalentes para VCR y solvente, con excepción de las muestras VCR 1cH. Asimismo, la producción de burbujas no afecta la CE en una medida capaz de distinguir las diferentes maneras de sucución. Concluimos que las propiedades físico-químicas de HD pueden ser modeladas por mecanismos químicos y difusivos típicos del agua destilada. Palabras clave: sucución, conductividad eléctrica, alta diluciones, agua destilada, modelo A maneira mais comum de realizar sucussões é manter uma preparação líquida dentro de um frasco de vidro, e agitá-la violentamente usando um dispositivo mecânico ou batendo-a, manualmente, contra um anteparo rígido, porém elástico. Este procedimento tem sido interpretado como um mecanismo capaz de transferir a energia mecânica da agitação, para níveis moleculares, onde ela se tornaria disponível para a realização de trabalho químico. Tal interpretação tem sido estimulada pela observação de alterações na condutividade elétrica (CE) de Altas Diluições (HD), devido à sucussão. Visando modelar esta questão, comparou-se a alteração da CE de amostras preparadas a partir de Sulfato de Vincristina (VCR) com amostras equivalentes (controle) preparadas apenas com o solvente água destilada. As amostras foram produzidas por sucussão mecânica e manual, a fimd e observar a influência da produção de bolhas. Os resultados confirma que a CE varia com o tempo, porém tais variações são equivalentes para VCR e controle, exceto para as amostras VCR 1cH. Também, a produção de bolhas não afeta a CE de forma a permitir usá-la para distinguir os diferentes procedimentos de sucussão. Concluimos que as propriedades físico-químicas da HDs podem ser modeladas por mecanismos químicos e difusivos, típicos para a água destilada.Palavras-Chave: sucussão, condutividade elétrca, altas diluições, água destilada, modelagem
    International Journal of High Dilution Research. 01/2008;
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    ABSTRACT: The latex of Euphorbia tirucalli L. (Aveloz) is popularly used to treat cancers from diverse sources such as: breast, prostate, lung and kidney. Furthermore, high dilutions of this latex (latex-HDs) have been successfully used in the treatment of tumors, although the mechanisms involved in this antitumoral activity are not yet known. The present study aimed to investigate the cytotoxic effects of latex-HDs in 2 human cell lines, including non-tumoral melanocytes (MelanA line) and human breast cancer cells (MCF7 line). Latex-HDs were obtained through the interaction of two procedures: 1:100 dilution in mass and agitation, using 2 solvents, water and 70% ethanol in homeopathic preparations 5, 15 and 30cH, following the Brazilian Homeopathic Pharmacopeia. Cell lines were incubated with 1 % and 10 % of latex-HDs for 24 hours. Controls treated with similar preparations without latex were incubated under the same conditions. Cell viability was analyzed by MTT assay and morphological features were assessed by May-Grunwald-Giemsa method. Cell ultrastructure was analyzed by transmission electron microscopy. Metabolic alterations were detected by spectrophotometric assay for the enzyme 6-phosphofructo-1-kinase (PFK-1) activity. 1% of ethanolic latex-HDs induced no alterations in MelanA cells viability, however Aveloz 15cH induced an increase in MCF7 cells viability. After aqueous treatment, MelanA cells viability decreased in almost all systems, whereas in MCF7 cells, an increase was detected in dynamized water systems and Aveloz 15CH. However, only water 15cH increased cell viability in comparison to control and non-dynamized water. No alterations in MCF7 morphology and ultrastructure were visualized through light microscopy or electronic microscopy, respectively. Interesting results were detected when we studied the glycolytic metabolism of both cells, giving support to evidences showing that HDs interfere in the metabolism of cell lines. Further studies are needed to better understand the mechanisms involved in this in vitro biological response to HDs. Keywords: Euphorbia tirucalli L.; Aveloz; PFK-1; High Dilutions; Cell metabolism. Altas diluições de Euphorbia tirucalli L. (aveloz) modificam a viabilidade e o metabolismo glicolítico de linhas celulares Resumo O látex de Ephorbia tirucalli L. (aveloz) é usado popularmente no tratamento de diversos tipos de câncer incluindo mama, próstata, pulmão e rim. Além do mais, altas diluições desse látex (latex-HDs) têm sido utilizadas com sucesso no tratamento de tumores, embora o mecanismo dessa ação ainda seja desconhecido. O presente trabalho objetivou pesquisar os efeitos citotóxicos de látex-HDs em 2 linhas celulares humanas, incluindo melanócitos não tumorais (línha MelanA) e células de câncer de mama humano (linha MCF7). As látex-HDs foram obtidas através de 2 procedimentos: diluição 1:100 em massa e agitação, com 2 solventes, água e etanol 70%, nas diluições homeopáticas 5, 15cH e 30cH de acordo com a Farmacopéia Homeopática Brasileira. As linhas celulares foram incubadas com látex-HDs 1% e 10% por 24 horas. Foram preparados controles similares sem o látex e incubados sob as mesmas condições. A viabilidade celular foi analisada através do método May-grunwald-Giemsa. A ultrastrutura celular foi analisada através de microscopia eletrônica de transmissão. As alterações metabólicas foram detectadas por ensaio de espectrofotometria para a atividade da anzima 6-fosfofructo-1-quinase (PFK-1). Látex-Hds em etanol 1% não induziram qualquer alteração na viabilidade das células MelanA, mas Aveloz 15cH induziu aumento da viabilidade das células MCF7. Após tratamento, a viabilidade das células MelanA diminuiu em quase todos os sistemas, enquanto que nas células MCF7 foi detectado aumento nos sistemas aquosos e Aveloz 15cH, mas só água 15cH aumentou a viabilidade celular por comparação ao controle e agua não dinamizada. Não foram detectadas alterações na morfologia celular por microscopis de luz quando se estudou o metabolismo glicolítico em ambas as células, demonstrando que os padrões de resposta celular dependem do tipo de solvente, metabolismo celular e procedimento de diluição. esa é a primeira vez que se mostra que as altas diluições interferem no metabolismo de linhas celulares. Outros estudos são necessários para se compreender melhor os mecanismos envolvidos nesta resposta in vitro a altas diluições de aveloz. Palavras-chave: Euphorbia tirucalli L.; Aveloz; PFK-1; Altas diluições; Metabolismo celular. Altas Diluciones de Euphorbia tirucalli L. (arbusto de goma) modifican la viabilidad y metabolismo glucolítico de líneas celulares Resumen El látex de Euphorbia tirucalli L. (arbusto de goma) es popularmente utilizado en el tratamiento de diversos tipos de cáncer, incluyendo mama, próstata, pulmón y riñón. Además, altas diluciones de este látex (latex-HDs) han sido utilizadas con éxito en el tratamiento de tumores, aunque el mecanismo de esta acción todavía es desconocido. Este trabajo tuvo por objetivo investigar los efectos citotóxicos de látex-HDs en 2 líneas celulares humanas, incluyendo melanocitos no tumorales (línea MelanA) y células de cáncer de mama humano (línea MCF7). Las latex-HDs fueron obtenidas mediante 2 procedimientos; dilución 1:100 en masa y agitación, con 2 solventes, agua y etanol 70%, en las diluciones homeopáticas 5, 15 y 30cH, según la Farmacopea Homeopática Brasileña. Las líneas celulares fueron incubadas con latex-HDs 1% y 10% durante 24 horas. Fueron preparados controles similares sin látex e incubados bajo las mismas condiciones. La viabilidad celular fue analizada por el método May-Grunwald-Giemsa. La ultra-estructura celular fue analizada por microscopía electrónica de transmisión. Alteraciones metabólicas fueron detectadas por ensayo de espectrofotometría para la actividad de la enzima 6-fosfofructo-1-kinasa (PFK-1). Látex-HDs en etanol 1% no indujeran ninguna alteración en la viabilidad de las células MelanA, sin embargo Euphorbia tirucalli 15cH indujo aumento de la viabilidad de las células MCF7. Después del tratamiento, la viabilidad de las células MelanA disminuyó en casi todos los sistemas, mientras que en las células MCF7 fue detectado aumento en los sistemas acuosos y Euphorbia tirucalli 15cH, pero solamente agua 15cH aumentó la viabilidad celular en comparación con el control y agua no dinamizada. No fueron detectadas alteraciones en la morfología celular por microscopía de luz cuando se estudió el metabolismo glucolítico de ambas células demostrando los modelos de respuesta celular dependen del tipo de solvente, metabolismo celular y procedimiento de dilución. Esta es la primera vez que se demostró que altas diluciones interfieren en el metabolismo de líneas celulares. Otros estudios son necesarios para entender mejor los mecanismos implicados en esta respuesta in vitro a las altas diluciones de Euphorbia tirucalli. Palabras-clave: Euphorbia tirucalli L.; Aveloz; PFK-1; Altas diluciones; metabolismo celular. Correspondence author: Carla Holandino Quaresma, cholandino@yahoo.com How to cite this article: Aquino CL, Barbosa GM, Barbosa GM, Varricchio MCBA, Veiga VF, Kuster R, Zancan P, Sola-Penna M, Quaresma CH. High dilutions of Euphorbia tirucalli L. (AVELOZ) modify the viability and glycolytic metabolism of cell lines. Int J High Dilution Res [online]. 2008 [cited YYYY Mmm DD]; 7(24): 132-139. Available from: http://www.feg.unesp.br/~ojs/index.php/ijhdr/article/view/283/361. El látex de Euphorbia tirucalli L. (arbusto de goma) es popularmente utilizado en el tratamiento de diversos tipos de cáncer, incluyendo mama, próstata, pulmón y riñón. Además, altas diluciones de este látex (latex-HDs) han sido utilizadas con éxito en el tratamiento de tumores, aunque el mecanismo de esta acción todavía es desconocido. Este trabajo tuvo por objetivo investigar los efectos citotóxicos de látex-HDs en 2 líneas celulares humanas, incluyendo melanocitos no tumorales (línea MelanA) y células de cáncer de mama humano (línea MCF7). Las latex-HDs fueron obtenidas mediante 2 procedimientos; dilución 1:100 en masa y agitación, con 2 solventes, agua y etanol 70%, en las diluciones homeopáticas 5, 15 y 30cH, según la Farmacopea Homeopática Brasileña. Las líneas celulares fueron incubadas con latex-HDs 1% y 10% durante 24 horas. Fueron preparados controles similares sin látex e incubados bajo las mismas condiciones. La viabilidad celular fue analizada por el método May-Grunwald-Giemsa. La ultra-estructura celular fue analizada por microscopía electrónica de transmisión. Alteraciones metabólicas fueron detectadas por ensayo de espectrofotometría para la actividad de la enzima 6-fosfofructo-1-kinasa (PFK-1). Látex-HDs en etanol 1% no indujeran ninguna alteración en la viabilidad de las células MelanA, sin embargo Euphorbia tirucalli 15cH indujo aumento de la viabilidad de las células MCF7. Después del tratamiento, la viabilidad de las células MelanA disminuyó en casi todos los sistemas, mientras que en las células MCF7 fue detectado aumento en los sistemas acuosos y Euphorbia tirucalli 15cH, pero solamente agua 15cH aumentó la viabilidad celular en comparación con el control y agua no dinamizada. No fueron detectadas alteraciones en la morfología celular por microscopía de luz cuando se estudió el metabolismo glucolítico de ambas células demostrando los modelos de respuesta celular dependen del tipo de solvente, metabolismo celular y procedimiento de dilución. Esta es la primera vez que se demostró que altas diluciones interfieren en el metabolismo de líneas celulares. Otros estudios son necesarios para entender mejor los mecanismos implicados en esta respuesta in vitro a las altas diluciones de Euphorbia tirucalli.Palabras-clave: Euphorbia tirucalli L.; Aveloz; PFK-1; Altas diluciones; metabolismo celular. O látex de Ephorbia tirucalli L. (aveloz) é usado popularmente no tratamento de diversos tipos de câncer incluindo mama, próstata, pulmão e rim. Além do mais, altas diluições desse látex (latex-HDs) têm sido utilizadas com sucesso no tratamento de tumores, embora o mecanismo dessa ação ainda seja desconhecido. O presente trabalho objetivou pesquisar os efeitos citotóxicos de látex-HDs em 2 linhas celulares humanas, incluindo melanócitos não tumorais (línha MelanA) e células de câncer de mama humano (linha MCF7). As látex-HDs foram obtidas através de 2 procedimentos: diluição 1:100 em massa e agitação, com 2 solventes, água e etanol 70%, nas diluições homeopáticas 5, 15cH e 30cH de acordo com a Farmacopéia Homeopática Brasileira. As linhas celulares foram incubadas com látex-HDs 1% e 10% por 24 horas. Foram preparados controles similares sem o látex e incubados sob as mesmas condições. A viabilidade celular foi analisada através do método May-grunwald-Giemsa. A ultrastrutura celular foi analisada através de microscopia eletrônica de transmissão. As alterações metabólicas foram detectadas por ensaio de espectrofotometria para a atividade da anzima 6-fosfofructo-1-quinase (PFK-1). Látex-Hds em etanol 1% não induziram qualquer alteração na viabilidade das células MelanA, mas Aveloz 15cH induziu aumento da viabilidade das células MCF7. Após tratamento, a viabilidade das células MelanA diminuiu em quase todos os sistemas, enquanto que nas células MCF7 foi detectado aumento nos sistemas aquosos e Aveloz 15cH, mas só água 15cH aumentou a viabilidade celular por comparação ao controle e agua não dinamizada. Não foram detectadas alterações na morfologia celular por microscopis de luz quando se estudou o metabolismo glicolítico em ambas as células, demonstrando que os padrões de resposta celular dependem do tipo de solvente, metabolismo celular e procedimento de diluição. esa é a primeira vez que se mostra que as altas diluições interferem no metabolismo de linhas celulares. Outros estudos são necessários para se compreender melhor os mecanismos envolvidos nesta resposta in vitro a altas diluições de aveloz.Palavras-chave: Euphorbia tirucalli L.; Aveloz; PFK-1; Altas diluições; Metabolismo celular.
    International Journal of High Dilution Research. 01/2008;
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    ABSTRACT: The latex extracted from Euphorbia tirucalli, a plant popularly known as Aveloz, is used in complementary medicine to induce tumor regression. However, as this latex has toxic effects when administered orally in ponderal doses, the present study was designed to assess the effects of high dilutions in healthy mice over a period of 18 weeks. The Aveloz latex-high diluted solutions (latex-HD) were obtained through the interaction of two processes: 1:100 dilution in mass and succussion, using ethanol 70% as a solvent, in the homeopathic dilutions of 5, 15 and 30cH, following Farmacopéia Homeopática Brasileira. Control solutions without latex were compounded (ethanolic-HD) in the same dilutions and were administered simultaneously. The animals which received latex-HD 30cH showed a significant increase in food consumption (p Keywords: Euphorbia tirucalli; Aveloz; Toxicology; High Dilutions; Mice. Efeitos toxicológicos crônicos de altas diluições de Aveloz (Euphorbia tirucalli L.) em camundongos sadios: estudo preliminar. Resumo O látex extraído de Euphorbia tirucalli, planta conhecida popularmente como Aveloz, é utilizado em medicina complementar para induzir regressão tumoral. No entanto, como este látex tem efeitos tóxicos quando administrado por via oral em doses ponderais, este estudo foi desenhado para avaliar os efeitos de altas diluições do mesmo em camundongos sadios por um período de 18 semanas. As altas diluições do látex do Aveloz (látex-HD) foram obtidas através da interação de dois processos: diluição 1:100 em massa e sucussão, utilizando etanol 70% como solvente, nas diluições homeopáticas 5, 15 e 30cH, de acordo com a Famracopéia Homeopática Brasileira. Soluções de controle sem o látex (etanol-HD) foram compostas nas mesmas diluições e ministradas simultaneamente. Os animais que receberam látex-HD 30cH mostraram aumento significativo no consumo de alimento (p Palavras-chave: Euphorbia tirucalli; Aveloz; Toxicologia; Altas diluições; Camundongos. Efectos toxicológicos crónicos de altas diluciones de Euphorbia tirucalli en ratones sanos: estudio preliminar. Resumen El látex extraído de Euphorbia tirucalli, planta conocida popularmente como arbusto de goma, es utilizado en medicina complementaria para la inducción de regresiones tumorales. Como este látex produce efectos tóxicos cuando es administrado por vía oral en dosis ponderales, este estudio fue diseñado para evaluar los efectos de altas diluciones de éste en ratones sanos, durante 18 semanas. Las altas diluciones del látex de E. tirucalli (látex-HD) fueron preparadas mediante la interacción de dos procesos: dilución 1:100 en masa y sucusión, utilizando etanol 70% como solvente en las diluciones homeopáticas 5, 15 y 30cH, según la Farmacopea Homeopática Brasileña. Soluciones de control, sin látex (etanol-HD) fueron compuestas en estas mismas diluciones y administradas simultáneamente. Los animales que recibieron látex-HD 30cH mostraron un aumento significativo del consumo de alimento (p Palabras-clave: Euphorbia tirucalli; Aveloz; Toxicologií; Altas diluciones; Ratones. Correspondence author: Carla Holandino Quaresma, cholandino@yahoo.com How to cite this article: Varricchio MCBN, Pereira C, Sales F, Gomes T, Daudt E, Aquino CL, Barbosa GM, Gomes N, Pyrrho AS, Hobaica PEM, Branco MC, Kuster R, Holandino C. Chronic toxicological effects of high diluted solutions of Aveloz (Euphorbia tirucalli L.) on healthy mice: a preliminary study. Int J High Dilution Res [online]. 2008 [cited YYYY Mmm DD]; 7(25): 174-178. Available from: http://www.feg.unesp.br/~ojs/index.php/ijhdr/article/view/308/371. O látex extraído de Euphorbia tirucalli, planta conhecida popularmente como Aveloz, é utilizado em medicina complementar para induzir regressão tumoral. No entanto, como este látex tem efeitos tóxicos quando administrado por via oral em doses ponderais, este estudo foi desenhado para avaliar os efeitos de altas diluições do mesmo em camundongos sadios por um período de 18 semanas. As altas diluições do látex do Aveloz (látex-HD) foram obtidas através da interação de dois processos: diluição 1:100 em massa e sucussão, utilizando etanol 70% como solvente, nas diluições homeopáticas 5, 15 e 30cH, de acordo com a Famracopéia Homeopática Brasileira. Soluções de controle sem o látex (etanol-HD) foram compostas nas mesmas diluições e ministradas simultaneamente. Os animais que receberam látex-HD 30cH mostraram aumento significativo no consumo de alimento (pPalavras-chave: Euphorbia tirucalli; Aveloz; Toxicologia; Altas diluições; Camundongos. El látex extraído de Euphorbia tirucalli, planta conocida popularmente como arbusto de goma, es utilizado en medicina complementaria para la inducción de regresiones tumorales. Como este látex produce efectos tóxicos cuando es administrado por vía oral en dosis ponderales, este estudio fue diseñado para evaluar los efectos de altas diluciones de éste en ratones sanos, durante 18 semanas. Las altas diluciones del látex de E. tirucalli (látex-HD) fueron preparadas mediante la interacción de dos procesos: dilución 1:100 en masa y sucusión, utilizando etanol 70% como solvente en las diluciones homeopáticas 5, 15 y 30cH, según la Farmacopea Homeopática Brasileña. Soluciones de control, sin látex (etanol-HD) fueron compuestas en estas mismas diluciones y administradas simultáneamente. Los animales que recibieron látex-HD 30cH mostraron un aumento significativo del consumo de alimento (pPalabras-clave: Euphorbia tirucalli; Aveloz; Toxicologií; Altas diluciones; Ratones.
    International Journal of High Dilution Research. 01/2008;

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